| 深海沉积物微生物宏基因组文库中IMP环水解酶的筛选及基因克隆 |
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| 引用本文： | 陈兴麟,赵晶,徐宪忠,曾润颖. 深海沉积物微生物宏基因组文库中IMP环水解酶的筛选及基因克隆[J]. 应用与环境生物学报, 2011, 0(6): 855-859 |
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| 作者姓名： | 陈兴麟 赵晶 徐宪忠 曾润颖 |
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| 作者单位： | 国家海洋局第三海洋研究所海洋生物遗传资源重点实验室;厦门大学海洋与环境学院海洋技术与工程系; |
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| 基金项目： | 中国大洋协会项目(No.DYXM-115-02-2-04)资助~~ |
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| 摘 要： | 为了解嘌呤合成途径的关键酶——IMP环水解酶的多样性,从深海沉积物样品中提取总DNA构建Cosmid宏基因组文库,从中筛选出一个具有IMP环水解酶活性的克隆(CAIMP1).通过基因步移的方法从CAIMP1的约35 kb的插入片段中扩增出长度为1 599 bp的完整的IMP环水解酶基因,该基因包含MGS和AICARFT_IMPCHas两个保守结构域.活性检测结果表明CAIMP1克隆的发酵液上清具有较强的IMP环水解酶活性.该酶与数据库中收录的IMP环水解酶的氨基酸序列同源性较低,系统发育分析也表明该酶与其它参照序列亲缘关系较远,表明该序列可能为一个新的IMP环水解酶基因.
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| 关 键 词： | 深海沉积物 宏基因组文库 IMP环水解酶 保守结构域 |
Screening and Cloning of IMP Cyclohydrolase from a Metagenomic Cosmid Library of a Deep-sea Sediment Sample |
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| CHEN Xinglin,ZHAO Jing,XU Xianzhong , ZENG Runying. Screening and Cloning of IMP Cyclohydrolase from a Metagenomic Cosmid Library of a Deep-sea Sediment Sample[J]. Chinese Journal of Applied and Environmental Biology, 2011, 0(6): 855-859 |
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| Authors: | CHEN Xinglin ZHAO Jing XU Xianzhong & ZENG Runying |
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| Affiliation: | CHEN Xinglin1,ZHAO Jing2,XU Xianzhong & ZENG Runying1(1Key Laboratory of Marine Biogenetic Resources,Third Institude Oceanography,State Oceanic Administration of China,Xiamen 361005,China)(2Marine and Environment College,Xiamen University,China) |
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| Abstract: | IMP cyclohydrolase is a key enzyme in the purine synthesis pathway.A clone(CAIMP1) producing IMP cyclohydrolase activity was isolated from a metagenomic library which was constructed from metagenomic DNA of a deep-sea sediment sample.The gene coding for the IMP cyclohydrolase with the size of 1 599 bp was subcloned from the 35 kb inserted fragment of CAIMP1 by gene walking.Two conserved domains,MGS and AICARFT_IMPCHas,was found in the gene.The CAIMP1 exhibited high activity towards 5-formaminoimidazole-4-ca... |
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| Keywords: | deep-sea sediment metagenomic library IMP cyclohydrolase conserved domain |
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